Disubstituted pyrazole compounds

ABSTRACT

The present invention provides a compound of Formula I.or a pharmaceutically acceptable salt thereof, and the use of compounds of Formula I for treating metabolic conditions, such as type 2 diabetes mellitus, heart failure, diabetic kidney disease, and non-alcoholic steatohepatitis.

The present invention relates to novel ketohexokinase (KHK) inhibitorcompounds, to pharmaceutical compositions comprising the compounds andto the use of the compounds for the treatment of certain conditions,such as type 2 diabetes mellitus (T2DM), heart failure, diabetic kidneydisease and non-alcoholic steatohepatitis (NASH).

KHK, also referred to as fructokinase, is the rate-limiting enzymeinvolved in fructose metabolism. It catalyses the phosphorylation offructose to fructose-1-phosphate (F1P), causing concomitant depletion ofcellular ATP levels. In contrast to glucose, fructose metabolism lacksfeedback inhibition and it triggers accumulation of downstreamintermediates involved in, for example, lipogenesis, gluconeogenesis andoxidative phosphorylation (Hannou, S. A., et al.; J. Clin. Invest.,128(2), 544-555, 2018). This has negative metabolic consequences whichare associated with a number of serious metabolic disorders.

KHK exists in two alternatively spliced isoforms consisting of KHK-C andKHK-A differing in exon 3. KHK-C is expressed primarily in the liver,kidney and intestine, whereas KHK-A is more ubiquitous. Mice deficientin both isoforms are fully protected from fructose-induced metabolicsyndrome. However, the adverse metabolic effects are exacerbated in micelacking KHK-A only (Ishimoto T, et al.; Proc. Natl. Acad. Sci. USA,109(11), 4320-4325, 2012).

Several epidemiologic and experimental studies have reported thatincreased consumption of fructose, and more precisely increased fructosemetabolism, may play an important role in the development of certaindisorders, including metabolic syndrome and in particular, in thedevelopment of T2DM (Softic et al.; J. Clin. Invest., 127(11),4059-4074, 2017), heart failure (Mirtschink, P., et al.; Eur. Heart J.,39, 2497-2505, 2018), diabetic kidney disease (Cirillo, P., et al.; J.Am. Soc. Nephrol., 20, 545-553, 2009) and NAFLD/NASH (Vos, M. B., etal.; Hepatology, 57, 2525-2531, 2013). Targeting inhibition of KHK isexpected to limit fructose metabolism and provide effective treatmentoptions for a number of metabolic disorders.

US 2017/0183328 A1 discloses substituted 3-azabicyclo[3.1.0]hexanes asKHK inhibitors. Recently published data shows that ketohexokinaseinhibitor PF-06835919 administered for 6 weeks reduces whole liver fatas measured by magnetic resonance imaging-proton density fat fraction insubjects with non-alcoholic fatty liver disease (J. Hepatology. EASLInternational Liver Congress Abstracts, Supplement No 1S Vol. 70, April2019).

Compounds containing carboxylic functional groups carry a riskassociated with the formation of acyl glucuronide metabolites (Vleet Vanet al., Toxicology Letters, 272 (2017) 1-7). Acyl glucuronidemetabolites are often unstable and may be chemically reactive leading tocovalent bonding with macromolecules and toxicity.

There is a need for alternate treatments for metabolic syndrome andassociated indications including T2DM, heart failure, diabetic kidneydisease and NASH. In particular, there is a need for compounds which arepotent inhibitors of KHK. There is a need for KHK inhibitor compoundshaving advantageous properties, for example, good oral bioavailabilityto support once daily dosing. Furthermore, there is a need for KHKinhibitor compounds which do not have a carboxylic acid moiety and lackthe ability to form acyl glucuronides.

Accordingly, in one embodiment, the present invention provides acompound of the Formula I:

whereinX is N, or C substituted with CN;R¹ is selected from: H,

R² and R³ are both H, or one is H and the other is OH;R⁴, R⁵, R⁶, R⁷ and R⁹ are independently H or CH₃;R⁸ is H, CH₃, CH₂CH₂OH, C(═O)CH₂NH₂, or C(═O)CH₃; andR¹⁰ is OH or NH₂;or a pharmaceutically acceptable salt thereof.

In a particular embodiment, R¹ is selected from: H,

or a pharmaceutically acceptable salt thereof.

In a particular embodiment, there is provided a compound of Formula Iwherein X is N or C substituted with CN;

R¹ is selected from:

R² is H and R³ is OH,or R² is OH and R³ is H,or R² and R³ are both H;or a pharmaceutically acceptable salt thereof.

In a particular embodiment, the compound is of Formula Ia:

or a pharmaceutically acceptable salt thereof.

In a particular embodiment, the compound is of Formula Ic:

or a pharmaceutically acceptable salt thereof.

In a particular embodiment, the compound is of Formula Ie:

or a pharmaceutically acceptable salt thereof.

In an embodiment, X is N.

In an embodiment, X is C substituted with CN.

In an embodiment, R¹ is

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In a particular embodiment, there is provided a succinate salt of

In a preferred embodiment, the succinate salt is the sesquisuccinatesalt.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In an embodiment, the compound of Formula I is:

or a pharmaceutically acceptable salt thereof.

In a particular embodiment, the compound is selected from:

-   2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone;-   [(2R)-1-[4-[1-(4-piperidyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;-   [(2R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;-   (2S,3R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol;-   2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-N-(2-hydroxyethyl)acetamide;-   2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-6-[1-(1-methyl-4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-[1-(2-hydroxyethyl)-4-piperidyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-[2-(dimethylamino)ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-(2-hydroxyethyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-4-[1-(4-piperidyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine;-   (2S,3R)-2-methyl-1-[4-[1-[(3R)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol;-   (2S,3R)-2-methyl-1-[4-[1-[(3S)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol;-   [(2R)-1-[4-[1-[(3R)-pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;-   6-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-[1-(2-aminoacetyl)-4-piperidyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-(1-acetyl-4-piperidyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-6-[1-[2-(4-methylpiperazin-1-yl)-2-oxo-ethyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-N,N-bis(2-hydroxyethyl)acetamide;-   6-[1-[2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-[2-[(3S,4S)-3,4-dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-6-(1H-pyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile;-   2-[(2S)-2-methylazetidin-1-yl]-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile;-   6-[1-(3-hydroxy-4-piperidyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile;-   N-(2-aminoethyl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetamide;-   2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(3S)-3-methylpiperazin-1-yl]ethanone;-   2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(2S)-2-methylpiperazin-1-yl]ethanone;-   1-(3,3-dimethylpiperazin-1-yl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]ethanone;-   1-[(2S,5R)-2,5-dimethylpiperazin-1-yl]-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]ethanone;-   4-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidine;-   2-[(2S)-2-methylazetidin-1-yl]-4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine;-   (2S,3R)-2-methyl-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol;    or-   [(2R)-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol;-   or a pharmaceutically acceptable salt thereof.

In a particular embodiment, the compound is2-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanonesesquisuccinate, which is also known as ethanone,2-[4-[2-[(2S)-2-methyl-1-azetidinyl]-6-(trifluoromethyl)-4-pyrimidinyl]-1H-pyrazol-1-yl]-1-(1-piperazinyl)-,butanedioate (1:1.5) or butanedioicacid-2-(4-{2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl}-1H-pyrazol-1-yl)-1-(piperazin-1-yl)ethan-1-one(1.5/1).

Formula I encompasses Formulae Ia, Ib, Ic, Id, Ie, If, Ig and Ih andreference to Formula I below, for example in the methods of treatmentand therapeutic uses, is also be read as a reference to each and all ofthese sub-formulae.

In an embodiment, there is provided a method of treating T2DM in apatient in need of such treatment, comprising administering to thepatient an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

In an embodiment, there is provided a method of treating heart failurein a patient in need of such treatment, comprising administering to thepatient an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

In an embodiment, there is provided a method of treating diabetic kidneydisease in a patient in need of such treatment, comprising administeringto the patient an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

In an embodiment, there is provided a method of treating NASH in apatient in need of such treatment, comprising administering to thepatient an effective amount of a compound of Formula I, or apharmaceutically acceptable salt thereof.

In an embodiment, there is provided a method of treating a diseaseselected from the group consisting of metabolic syndrome, NAFLD,obesity, cardiovascular disease, coronary artery disease, chronic kidneydisease, dyslipidermia and diabetic complications for example diabeticretinopathy, in a patient in need of such treatment, comprisingadministering to the patient an effective amount of a compound ofFormula I, or a pharmaceutically acceptable salt thereof.

Furthermore, in an embodiment, there is provided a compound of FormulaI, or a pharmaceutically acceptable salt thereof, for use in therapy. Inan embodiment, there is provided a compound of Formula I, or apharmaceutically acceptable salt thereof, for use in treating T2DM. Inan embodiment, there is provided a compound of Formula I, or apharmaceutically acceptable salt thereof, for use in treating heartfailure. In an embodiment, there is provided a compound of Formula I, ora pharmaceutically acceptable salt thereof, for use in treating diabetickidney disease. In an embodiment, there is provided a compound ofFormula I, or a pharmaceutically acceptable salt thereof, for use intreating NASH. In an embodiment, there is provided a compound of FormulaI, or a pharmaceutically acceptable salt thereof, for use in treatingmetabolic syndrome, NAFLD, obesity, cardiovascular disease, coronaryartery disease, chronic kidney disease, dyslipidemia or diabeticcomplications for example diabetic retinopathy.

Furthermore, in an embodiment, there is provided the use of a compoundof Formula I, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for treating T2DM. In an embodiment, thereis provided the use of a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, for the manufacture of a medicament fortreating heart failure. In an embodiment, there is provided the use of acompound of Formula I, or a pharmaceutically acceptable salt thereof,for the manufacture of a medicament for treating diabetic kidneydisease. In an embodiment, there is provided the use of a compound ofFormula I, or a pharmaceutically acceptable salt thereof, for themanufacture of a medicament for treating NASH. In an embodiment, thereis provided the use of a compound of Formula I, or a pharmaceuticallyacceptable salt thereof, for the manufacture of a medicament fortreating metabolic syndrome, NAFLD, obesity, cardiovascular disease,coronary artery disease, chronic kidney disease, dyslipidemia ordiabetic complications for example diabetic retinopathy.

In an embodiment, there is provided a pharmaceutical composition,comprising a compound of Formula I, or a pharmaceutically acceptablesalt thereof, with one or more pharmaceutically acceptable carriers,diluents, or excipients. In an embodiment, there is provided a processfor preparing a pharmaceutical composition, comprising admixing acompound of Formula I, or a pharmaceutically acceptable salt thereof,with one or more pharmaceutically acceptable carriers, diluents, orexcipients.

As used herein, the terms “treating” or “to treat” includes restraining,slowing, stopping, or reversing the progression or severity of anexisting symptom or disorder.

As used herein, the term “patient” refers to a mammal. Preferably, thepatient is human.

As used herein, the term “effective amount” refers to the amount or doseof compound of Formula I, or a pharmaceutically acceptable salt thereof,which, upon single or multiple dose administration to the patient,provides the desired effect in the patient under diagnosis or treatment.

An effective amount can be determined by one skilled in the art by theuse of known techniques and by observing results obtained underanalogous circumstances. In determining the effective amount for apatient, a number of factors are considered, including, but not limitedto: the species of patient; its size, age, and general health; thespecific disease or disorder involved; the degree of or involvement orthe severity of the disease or disorder; the response of the individualpatient; the particular compound administered; the mode ofadministration; the bioavailability characteristics of the preparationadministered; the dose regimen selected; the use of concomitantmedication; and other relevant circumstances. The compounds of Formula Iare effective at a dosage per day that falls within the range of about0.1 to about 15 mg/kg of body weight.

The compounds of Formula I are formulated as pharmaceutical compositionsadministered by any route which makes the compound bioavailable.Preferably, such compositions are for oral administration. Suchpharmaceutical compositions and processes for preparing same are wellknown in the art (See, e.g., Remington, J. P., “Remington: The Scienceand Practice of Pharmacy”, L. V. Allen, Editor, 22^(nd) Edition,Pharmaceutical Press, 2012).

The compounds of Formula I and the pharmaceutically acceptable saltsthereof may be used in the methods of treatment and therapeutic uses ofthe invention, with certain configurations being preferred. It will beunderstood that the following preferences are applicable both to thetreatment methods, the therapeutic uses and to the compounds of theinvention.

Compounds of the present invention include:

and pharmaceutically acceptable salts thereof.

Although the present invention contemplates all individual enantiomersand diasteromers, as well as mixtures of said compounds, includingracemates, compounds of Formula Ia, Formula Ic and Formula Ie, andpharmaceutically acceptable salts thereof, are particularly preferred.

Individual enantiomers may be separated or resolved by one of ordinaryskill in the art at any convenient point in the synthesis of compoundsof Formula I, by methods such as selective crystallization techniques,chiral chromatography (See for example, J. Jacques, et al.,“Enantiomers, Racemates, and Resolutions”, John Wiley and Sons, Inc.,1981, and E. L. Eliel and S. H. Wilen, “Stereochemistry of OrganicCompounds”, Wiley-Interscience, 1994), or supercritical fluidchromatography (SFC) (See for example, T. A. Berger; “SupercriticalFluid Chromatography Primer,” Agilent Technologies, July 2015).

A pharmaceutically acceptable salt of the compounds of Formula I can beformed, for example, by reaction of an appropriate free base of acompound of Formula I and an appropriate pharmaceutically acceptableacid in a suitable solvent under standard conditions well known in theart (See, for example, Bastin, R. J., et al.; Org. Process. Res. Dev.,4, 427-435, 2000 and Berge, S. M., et al.; J. Pharm. Sci., 66, 1-19,1977). A preferred salt is a succinate salt. A particularly preferredsalt is the sesquisuccinate salt. In the sesquisuccinate salt, the ratioof free base:succinate is 1:1.5. The succinate salt is also known as thebutanedioate salt.

The compounds of Formula I, or salts thereof, may be prepared by avariety of procedures known to one of ordinary skill in the art, some ofwhich are illustrated in the schemes, preparations, and examples below.The products of each step in the schemes below can be recovered byconventional methods well known in the art, including extraction,evaporation, precipitation, chromatography, filtration, trituration, andcrystallization. In the schemes below, all substituents unless otherwiseindicated, are as previously defined. The reagents and startingmaterials are readily available to one of ordinary skill in the art.

Without limiting the scope of the invention, the following schemes,preparations, and examples are provided to further illustrate theinvention. In addition, one of ordinary skill in the art appreciatesthat compounds of Formula I may be prepared by using starting materialor intermediate with the corresponding desired stereochemicalconfiguration which can be prepared by one of skill in the art.

Certain abbreviations are defined as follows: “ACN” refers toacetonitrile; “BOC” refers to tert-butoxycarbonyl; “DCM” refers tomethylene chloride or dichloromethane; “DIPEA” refers toN,N-diisopropylethylamine; “DMF” refers to N,N-dimethylformamide; “DMSO”refers to dimethyl sulfoxide; “ELSD” refers to Evaporative lightscattering detector; “ES/MS” refers to Electrospray Mass Spectrometry;“EtOAc” refers to ethyl acetate; “EtOH” refers to ethanol or ethylalcohol; “h” refers to hour or hours; “HATU” refers to1-[Bis(dimethyl-amino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate; “HPLC” refers to high-performance liquidchromatography; “IPA” refers to isopropyl alcohol; “Me” refers tomethyl; “MeOH” refers to methanol; “MTBE” refers to methyl-tert-butylether; “min” refers to minute or minutes; “m/z” refers to mass-to-chargeratio; “Ph” refers to phenyl; “RBF” refers to round bottom flask; “RT”refers to room temperature; “SCX” refers to selective cation exchange;“SEM” refers to standard error of the mean; “SFC” refers tosupercritical fluid chromatography; “TFA” refers to trifluoroaceticacid; “THF” refers to tetrahydrofuran.

Scheme 1 depicts the general preparation of the compounds of Formula I.R^(1a) may be the same as R¹ in the final compound of Formula I or itmay be a group requiring transformation to reach R¹ of Formula I. Theroutine synthetic transformations of R^(1a), such as BOC-deprotection,ester hydrolysis and amide coupling reactions may be performed eitherbefore or after Step B. In Step A, a Suzuki cross-coupling reactionbetween heteroaryl dichloride 1 and pyrazole boronate ester 2 yieldssubstituted pyrazole compound 3. This reaction is performed using abase, for example 2 M aqeuous Na₂CO₃, in an organic solvent, for example1,4-dioxane, in the presence of a palladium catalyst, such astetrakis(triphenylphosphine)palladium(0) orbis(triphenylphosphine)palladium(II) dichloride, at elevatedtemperature. In Step B, compound 3 is subjected to a nucleophilicaromatic substitution reaction with substituted azetidine 4 to give acompound of Formula I. This reaction is performed in the presence of anorganic base, such as DIPEA, in an organic solvent, such as 1,4-dioxane,at elevated temperature.

Scheme 2 depicts the preparation of a subset of the compounds of FormulaI wherein the pyrazole is substituted with a nitrogen-containingheterocycle (Formula I′). Pyrazole boronate ester 5 is substituted witha nitrogen containing heterocycle as depicted in Scheme 2, wherein “Y”and “Z” may each independently be —CH₂— or —CH₂—CH₂—, and “PG” is anitrogen protecting group such as BOC. In Step C, a Suzukicross-coupling reaction between heteroaryl dichloride 1 and pyrazoleboronate ester 5 yields substituted pyrazole compound 6. As described inStep A of Scheme 1, this reaction is performed using a base, for example2 M aqeuous Na₂CO₃, in an organic solvent, for example 1,4-dioxane, inthe presence of a palladium catalyst, such astetrakis(triphenylphosphine)palladium(0) orbis(triphenylphosphine)palladium(II) dichloride, at elevatedtemperature. Two different routes can be taken as depicted in Scheme 2,Route A and Route B.

In Route A, substituted pyrazole compound 6 undergoes deprotection inStep D to give compound 7. For example if “PG” is BOC, deprotection canbe accomplished with TFA. In Step E, the heterocyclic nitrogen ofcompound 7 can undergo substitution reactions, for example reductiveamination, acylation, or amide coupling to give compound 8. Suchsubstitutions are depicted as “R” in this scheme. In Step F, compound 8is subjected to a nucleophilic aromatic substitution reaction withsubstituted azetidine 4 to give compound of Formula I′. As described inStep B of Scheme 1, this reaction is performed in the presence of anorganic base, such as DIPEA, in an organic solvent, such as 1,4-dioxane,at elevated temperature. This may be the final step or alternatively,the R group may be subjected to further routine synthetictransformations such as protecting group removal.

In Route B, substituted pyrazole compound 6 undergoes nucleophilicaromatic substitution reaction in Step G with substituted azetidine 4 togive compound 9. This can be accomplished by nucleophilic aromaticsubstitution reaction with substituted azetidine 4 in the presence of anorganic base, such as DIPEA, in an organic solvent, such as 1,4-dioxaneat elevated temperature. Alternatively, Steps C and G can beaccomplished in a one-pot procedure, wherein the Suzuki cross-couplingreaction of Step C is completed first, and then azetidine 4 is added tothe reaction along with an organic base (e.g. DIPEA), and then Step Gproceeds at elevated temperature. Protecting group “PG” is removed inStep H (e.g. with TFA if PG is BOC) to give compound 10. In Step I thenitrogen group of compound 10 can undergo substitution reactions, forexample reductive amination, acylation, or amide coupling to givecompound of Formula I′. This may be the final step or alternatively, theR group may be subjected to further routine synthetic transformationssuch as protecting group removal.

Scheme 3 depicts the preparation of a subset of the compounds of FormulaI wherein the pyrazole is substituted with an acetamide group (FormulaI″). In Step J, a Suzuki cross-coupling reaction between heteroaryldichloride 1 and pyrazole boronate ester 11 yields substituted pyrazolecompound 12. As described in Schemes 1 and 2, this reaction is performedusing a base, for example 2 M aqeuous Na₂CO₃, in an organic solvent, forexample 1,4-dioxane, in the presence of a palladium catalyst, such astetrakis(triphenylphosphine)palladium(0) orbis(triphenylphosphine)palladium(II) dichloride, at elevatedtemperature. In Step K, heteroaryl chloride 12 undergoes nucleophilicaromatic substitution with azetidine 4 in the presence of an organicbase, such as DIPEA, in an organic solvent, such as 1,4-dioxane, atelevated temperature to give compound 13. In Step L, ester hydrolysisusing a base e.g. sodium hydroxide gives acid 14. In Step M, acid 14undergoes amide coupling reaction with an amine of the formula HNR′R″ togive amide compound of Formula I″. The amine HNR′R″ can be cyclic (e.g.an optionally substituted piperazine). Step M may be the final step orthere may be further routine synthetic transformations such asprotecting group removal.

PREPARATIONS AND EXAMPLES

The following Preparations and Examples further illustrate variousembodiments of the invention and represent typical synthesis of thecompounds of the invention. The reagents and starting materials arereadily available or may be readily synthesized by one of ordinary skillin the art. It should be understood that the Preparations and Examplesare set forth by way of illustration and not limitation, and thatvarious modifications may be made by one of ordinary skill in the art.

LC-ES/MS is performed on an AGILENT® HP1200 liquid chromatographysystem. Electrospray mass spectrometry measurements (acquired inpositive and/or negative mode) are performed on a Mass SelectiveDetector quadrupole mass spectrometer interfaced to an HPLC which may ormay not have an ELSD. LC-MS conditions (low pH): column: PHENOMENEX®GEMINI® NX C18 2.0×50 mm 3.0 μm, 110 Å; gradient: 5-95% B in 1.5 min,then 95% B for 0.5 min column temperature: 50° C.+/−10° C.; flow rate:1.2 mL/min; 1 μL injection volume; Solvent A: deionized water with 0.1%HCOOH; Solvent B: ACN with 0.1% formic acid; wavelength 200-400 nm and212-216 nm. If the HPLC is equipped with an ELSD the settings are 45° C.evaporator temperature, 40° C. nebulizer temperature, and 1.6 SLM gasflow rate. Alternate LC-MS conditions (high pH): column: Waters xBridge®C18 column 2.1×50 mm, 3.5 μm; gradient: 5-95% B in 1.5 min, then 95% Bfor 0.50 min; column temperature: 50° C.+/−10° C.; flow rate: 1.2mL/min; 1 μL injection volume; Solvent A: 10 mM NH₄HCO₃ pH 9; Solvent B:ACN; wavelength: 200-400 nm and 212-216 nm; if had ELSD: 45° C.evaporator temp, 40° C. nebulizer temp, and 1.60 SLM gas flow rate.

The XRPD patterns of crystalline solids are obtained on a Bruker D4Endeavor X-ray powder diffractometer, equipped with a CuKα, source and aVantec detector, operating at 35 kV and 50 mA. The sample is scannedbetween 4 and 40 2θ°, with a step size of 0.008 2θ° and a scan rate of0.5 seconds/step, and using 1.0 mm divergence, 6.6 mm fixedanti-scatter, and 11.3 mm detector slits. The dry powder is packed on aquartz sample holder and a smooth surface is obtained using a glassslide. The crystal form diffraction patterns are collected at ambienttemperature and relative humidity. Crystal peak positions are determinedin MDI-Jade after whole pattern shifting based on an internal NIST 675standard with peaks at 8.853 and 26.774 2θ°. It is well known in thecrystallography art that, for any given crystal form, the relativeintensities of the diffraction peaks may vary due to preferredorientation resulting from factors such as crystal morphology and habit.Where the effects of preferred orientation are present, peak intensitiesare altered, but the characteristic peak positions of the polymorph areunchanged. See, e.g. The United States Pharmacopeia #23, NationalFormulary #18, pages 1843-1844, 1995. Furthermore, it is also well knownin the crystallography art that for any given crystal form the angularpeak positions may vary slightly. For example, peak positions can shiftdue to a variation in the temperature at which a sample is analyzed,sample displacement, or the presence or absence of an internal standard.In the present case, a peak position variability of ±0.2 2θ is presumedto take into account these potential variations without hindering theunequivocal identification of the indicated crystal form. Confirmationof a crystal form may be made based on any unique combination ofdistinguishing peaks.

Preparation 1 (2S)-1-Benzhydryl-2-methyl-azetidine[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid salt

Assemble a 2000 mL 3-neck RBF with an addition funnel, nitrogen inletand a thermometer adapter. Purge the vessel with nitrogen and add(3R)-butane-1,3-diol (25 g, 277 mmol), DIPEA (127 mL, 731 mmol) and ACN(556 mL). Cool the mixture to −30° C. Add trifluoromethanesulfonicanhydride (101 mL, 601 mmol) dropwise over 3 h such that the internaltemperature is maintained between −35 and −30° C. After the completionof the addition, stir for 10 min at −35 to −30° C. Addtrifluoromethanesulfonic anhydride (1.9 mL, 11 mmol) dropwise over 5 minsuch that the internal temperature is maintained between −35 and −30° C.After the completion of the addition, stir for 10 min at −35 to −30° C.Add DIPEA (127 mL, 731 mmol) dropwise over 15 min such that the internaltemperature is maintained between −35 and −30° C. After the completionof the addition, stir for 10 min at −35 to −30° C. In a separate flaskunder nitrogen, dissolve aminodiphenylmethane (48.0 mL, 270 mmol) in ACN(49 mL) and transfer the resulting solution to the addition funnel. Addthe amine solution to the cold triflate dropwise over 40 min such thatthe internal temperature is maintained between −20 to −35° C. After thecompletion of the addition, stir for 30 min at −35 to −30° C. Transferthe reaction to a water bath and allow it to slowly warm over 30 min.Remove the bath and allow the reaction to warm to RT over 30 min.Transfer the vessel to a heating mantle and warm the reaction to 45° C.for 30 min, then cool to RT. Pour the resulting mixture into 1200 mL ofwater and extract with toluene (400 mL×3). Combine the extracts and washwith water and saturated aqueous NaCl solution. Dry the organics overanhydrous Na₂SO₄, filter and concentrate in vacuo. Dry the residue undervacuum overnight, then dissolve it in DCM (400 mL). Prepare a silica gelpad on a fritted funnel and equilibrate it with 1:1 heptane/EtOAc. Loadthe product solution onto the silica gel pad and wash with 1600 mL of1:1 heptane/EtOAc. Concentrate the filtrate to give a red oil. Dissolvethe oil in MeOH (250 mL) and place the flask in a water bath (˜10° C.).Add L(−)-camphorsulfonic acid (61.6 g, 265 mmol) portion-wise keepingthe internal temperature below 20° C. Stir the resulting mixture for 15min and then concentrate in vacuo to give a brown foam. Dry the foam ona vacuum pump for 2 h. Dissolve the foam in DCM (130 mL), then slowlyadd EtOAc (1100 mL) to the stirring solution via addition funnel.Transfer the resulting mixture to a 4000 mL beaker and stir open to theatmosphere overnight. Cool the beaker in an ice bath for 10 min. Collectthe precipitate in a fritted funnel by vacuum filtration washing with aminimal amount of ice-cold EtOAc. Dry the solid on the frit for 2 h.Dissolve the resulting white solid in a minimal amount of DCM, transferto a 2000 mL beaker and then dilute slowly with EtOAc until the clearsolution starts to become cloudy. Stir the suspension for 4 h while opento the atmosphere. Collect the solids by vacuum filtration using afritted funnel and dry on the frit overnight to give the title compound(111.8 g, 238.06 mmol, 86% Yield) as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 10.54-10.47 (m, 1H), 7.61 (d, J=7.3 Hz, 5H), 7.47-7.37 (m,7H), 5.85 (d, J=10.3 Hz, 1H), 4.68-4.61 (m, 1H), 3.91-3.83 (m, 2H), 3.37(s, 8H), 2.99 (d, J=14.6 Hz, 1H), 2.77-2.68 (m, 1H), 2.51-2.44 (m, 4H),2.30-2.16 (m, 2H), 1.91-1.81 (m, 2H), 1.42-1.28 (m, 3H), 1.08 (s, 3H),1.01 (d, J=6.6 Hz, 3H), 0.77 (s, 4H); >98% ee [HPLC: Chiralcel® OJ (10cm×4.6 mm, 5 μm), 5 mL/min, 40° C. isocratic 10% EtOH (0.2%^(i)PrNH₂)/CO₂].

Preparation 2[(1R,4S)-7,7-Dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt

To a 2250 mL Parr vessel add 20 wt % Pd(OH)₂ on carbon (6.62 g). Purgethe bottle with nitrogen and add MeOH (250 mL). To the resultingsuspension, slowly add (2S)-1-benzhydryl-2-methyl-azetidine[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid salt(111 g, 236 mmol) dissolved in MeOH (250 mL). Seal the vessel. Purgewith nitrogen followed by hydrogen and pressurize to 60 PSI. Vigorouslyshake the reaction vessel in a Parr Shaker apparatus for 15 h at RT.Purge the vessel with nitrogen and then filter the reaction mixturethrough a pad of Celite®, washing with MeOH. Concentrate the filtrate togive a white solid and dry under vacuum. Suspend the solid in 780 mL of1:1 MTBE/EtOAc and heat the mixture to 65° C. for 20 h then cool to RTand stir overnight. Collect the solids by filtration. Suspend the solidsin 380 mL of MTBE and stir at RT for 24 h. Collect the white solid byfiltration to give the title compound (41.5 g, 137 mmol, 58% Yield). ¹HNMR (400 MHz, DMSO-d₆) δ 8.68-8.55 (m, 1H), 4.51-4.42 (m, 1H), 3.91-3.75(m, 1H), 3.36 (s, 3H), 2.91 (d, J=14.6 Hz, 1H), 2.69-2.61 (m, 1H),2.52-2.46 (m, 2H), 2.28-2.22 (m, 1H), 2.17-2.10 (m, 1H), 1.96 (t, J=4.5Hz, 1H), 1.89-1.79 (m, 1H), 1.43 (d, J=6.7 Hz, 2H), 1.36-1.26 (m, 1H),1.05 (s, 2H), 0.75 (s, 2H).

Preparation 3 (R)-2-Azetidinemethanol hydrochloride

To a 2-neck RBF, equipped with a nitrogen inlet, add:(R)-1-(tert-butoxycarbonyl)azetidine-2-carboxylic acid (30 g, 146 mmol),THF (300 mL), and 4-methylmorpholine (17.7 mL, 161 mmol). Cool themixture to −10° C. and add isobutyl chloroformate (21 mL, 161 mmol)dropwise. Stir the mixture for 30 min and then warm to RT. Remove theresulting solid by filtration. Cool the filtrate to 0° C. and add asolution of sodium borohydride (11.1 g, 292 mmol) in water (90 mL)dropwise (caution: gas evolution). After the addition, warm the mixtureto RT and stir for 30 min. Dilute the mixture with MTBE (300 mL) andwater (100 mL). Wash the mixture with saturated aqueous NaHCO₃ (200 mL)and then saturated aqueous NaCl (200 mL). Dry the organic phase overMgSO₄, filter, and concentrate to dryness to give an oil (27 g).Carefully add HCl (4.0 M) in 1,4-dioxane (110 mL) [Caution: Gasevolution] and stir the resulting mixture for 3 h at RT. Evaporate thesolvent in vacuo to give the title compound as an oil (16 g, 89%). Usethis material directly in preparations 9, 11, 16 and 36.

Preparation 4 (2S,3R)-2-methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid

Step 1

Equip a 3-neck 500 mL RBF with an addition funnel and temperature probe.To the flask add but-2-en-1-ol (cis trans mixture) (23.7 mL, 267 mmol)and chloroform (200 mL). Cool the solution in an ice bath until theinternal temperature reaches 1.2° C. Add bromine (13.7 mL, 267 mmol)dropwise by addition funnel over 2 h at a rate of ˜1 drop/6 s. After theaddition is complete warm the reaction to RT and stir for 30 min. Turnoff the stirring and allow the reaction to stand for 3 days. Quench withsat. Na₂S₂O₃ solution and stir vigorously for 10 min. Allow the mixtureto stand for 3 days. Remove the organic layer and extract the aqueousfraction with DCM (×3). Combine the organics and dry over Na₂SO₄, filterand concentrate in vacuo to give 2,3-dibromobutan-1-ol (62.3 g, 269mmol).

Step 2 Procedure A

Equip a 3-neck 1 L RBF with an addition funnel and temperature probe. Tothe flask add 2,3-dibromobutan-1-ol (62.3 g, 269 mmol) and THE (180 mL).Place the flask in a water bath at RT. Add a solution of KOH (15.1 g,269 mmol) in water (135 mL) dropwise over 10 min via the additionfunnel. Stir at RT for 2 h. Separate the organic phase. Extract theaqueous fraction with 3×150 mL EtOAc. Combine the organics, wash oncewith ˜200 mL brine, dry over Na₂SO₄ and filter. Carefully concentratethe organics (100 mbar, 30° C. until minimum volume, then 10 mbar, 30°C. for 10 min) to give 37 g of a mixture of 2-(1-bromoethyl)oxirane (60%mass), 2,3-dibromobutan-1-ol (36% mass), and EtOAc (4% mass) asdetermined by ¹H NMR. To the mixture, add EtOH (100 mL),aminodiphenylmethane (36 mL, 208.6 mmol), and NaHCO₃ (26 g, 309 mmol).Heat the reaction mixture to 65° C. overnight. Cool to RT.

Procedure B

In a first flask, add 2,3-dibromobutan-1-ol (72.6 g, 313 mmol) and THE(200 mL). Place the flask in a water bath at RT. Add a solution of KOH(17.6 g, 314 mmol) in water (150 mL). Stir at RT overnight. Separate theorganic phase. Extract the aqueous fraction with EtOAc (3×150 mL).Combine the organics, wash once with ˜200 mL saturated aqueous NaClsolution, dry over Na₂SO₄ and filter. Concentrate the organic layer invacuo (100 mbar, 30° C. until minimum volume, then 10 mbar, 30° C. for10 min) to give 41.1 g of a mixture of 2-(1-bromoethyl)oxirane (75%mass), 2,3-dibromobutan-1-ol (22% mass), and EtOAc (3% mass) asdetermined by ¹H NMR.

In a second flask, add 2,3-dibromobutan-1-ol (10 g, 43 mmol) and THE (30mL). Place the flask in a water bath at RT. Add a solution of KOH (2.42g, 43.1 mmol) in water (20 mL). Stir at RT overnight. Separate theorganic phase. Extract the aqueous fraction with 3×50 mL EtOAc. Combinethe organics, wash once with ˜200 mL saturated aqueous NaCl solution,dry over Na₂SO₄ and filter. Concentrate the organic layer in vacuo (100mbar, 30° C. until minimum volume, then 10 mbar, 30° C. for 10 min) togive 5.4 g of a mixture of 2-(1-bromoethyl)oxirane (66% mass),2,3-dibromobutan-1-ol (32% mass), and EtOAc (2% mass) as determined by¹H NMR.

In a third flask, add 2,3-dibromobutan-1-ol (10 g, 43 mmol) and THE (30mL). Place the flask in a water bath at RT. Add a solution of KOH (2.42g, 43.1 mmol) in water (20 mL). Heat the mixture to 50° C. for 2 h. Coolto RT. Separate the organic phase. Extract the aqueous fraction with3×50 mL EtOAc. Combine the organics, wash once with ˜200 mL brine, dryover Na₂SO₄ and filter. Concentrate the organic layer in vacuo (100mbar, 30° C. until minimum volume, then 10 mbar, 30° C. for 10 min) togive 5.4 g of a mixture of 2-(1-bromoethyl)oxirane (70% mass),2,3-dibromobutan-1-ol (28% mass), and EtOAc (2% mass) as determined by¹H NMR.

Combine the mixtures from the three reactions together in a RBF to give51.9 g of 2-(1-bromoethyl)oxirane (73% mass). Add EtOH (100 mL),aminodiphenylmethane (44 mL, 255.0 mmol), and NaHCO₃ (32 g, 380.926mmol). Stir the resulting mixture at RT for 2 h and then heat to 65° C.and continue stirring overnight. Cool to RT. Combine the crude reactionmixtures from Procedures A and B together for purification. Remove thesolids by filtration, washing with EtOH. Concentrate the filtrate todryness. Dissolve the resulting oil in DCM. Wash the resulting solutiontwice with NH₄Cl solution, dry over Na₂SO₄, filter and concentrate to avolume of ˜150 mL. Allow the mixture to stand overnight. Remove thesolids by filtration. Purify the filtrate by normal phase silicachromatography (70% MTBE:hexanes) to provide crude1-benzhydryl-2-methyl-azetidin-3-ol (66.8 g). ES/MS (m/z): 254 (M+H).

Step 3

Dissolve 1-benzhydryl-2-methyl-azetidin-3-ol (66.8 g) in MeOH (608 mL).Purify the resulting solution using a 5×25 cm Lux i-Cellulose 5 columnusing a solvent system of 85/15 CO₂/EtOH with 0.5% dimethylethylamineand a flow rate of 300 mL/min to give(2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol (19.2 g).

Step 4

Charge a RBF with (2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol (19.2 g,75.8 mmol), L(−)-camphorsulfonic acid (19 g, 80.2 mmol), EtOH (100 mL)and Et₂O (50 mL). Heat the mixture until nearly all the solids dissolve,then briefly sonicate. Heat the mixture to reflux, then cool to RT andstore in the freezer overnight. Collect the solids by filtration, washwith a large volume of Et₂O and dry under reduced pressure to give(2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol;[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (31.9 g,65.7 mmol).

Step 5

Charge a RBF with 20% palladium hydroxide on carbon (50 wt-% water) (2g). Wet the catalyst with a small volume of EtOH. To the suspension adda partial solution of (2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol;[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (20 g,41.2 mmol) in EtOH (400 mL). Sparge the suspension with nitrogen for 5min and then briefly with hydrogen. Stir the reaction mixture under aballoon of hydrogen until all starting material is gone by LC-MSanalysis. Filter the reaction mixture through a pad of Celite®.

In a second flask, using the same procedure, hydrogenate(2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol;[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (10 g,20.6 mmol) with 20% palladium hydroxide on carbon (50 wt-% water) (1 g)in EtOH (200 mL).

In a third flask using the same procedure, hydrogenate(2S,3R)-1-benzhydryl-2-methyl-azetidin-3-ol;[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (2 g,4.12 mmol) with 20% Palladium hydroxide on carbon (50 wt-% water) (0.2g) in EtOH (40 mL).

Combine the three filtrates together and concentrate in vacuo. Suspendthe material in n-heptane and sonicate for 10 min and then collect thesolids by filtration. Repeat the sonication-filtration sequence fourmore times. Dry the solid under vacuum overnight to give the titlecompound (20.11 g; 53% yield over 5 steps). ¹H NMR (399.85 MHz, MeOD):4.35-4.25 (m, 2H), 4.07 (dd, J=6.9, 10.6 Hz, 1H), 3.78 (dd, J=7.0, 10.6Hz, 1H), 3.30 (m, 1H), 2.79 (d, J=14.9 Hz, 1H), 2.70-2.63 (m, 1H),2.40-2.33 (m, 1H), 2.08-2.03 (m, 2H), 1.92 (d, J=18.4 Hz, 1H), 1.69-1.62(m, 1H), 1.54 (d, J=6.6 Hz, 3H), 1.47-1.41 (m, 1H), 1.14 (s, 3H), 0.88(s, 3H).

Preparation 5 Ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate

Charge a 500 mL RBF with4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (15 g, 77.32mmol), ACN (150 mL), potassium carbonate (32.06 g, 232.0 mmol) and ethylbromoacetate (9.09 mL, 81.2 mmol). Stir the slurry at RT overnight.Partition the reaction mixture between EtOAc (300 mL) and water (250mL). Separate the layers and extract the aqueous layer with EtOAc.Combine the organics and wash with saturated aqueous sodium chloride(500 mL), dry over sodium sulfate, filter and concentrate in-vacuo togive a light yellow oil. Purify by silica gel chromatography using agradient of 0 to 5% MeOH in DCM to give the title compound (16.8 g,76%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.94 (s, 1H), 7.60 (s, 1H), 5.08 (s,2H), 4.14 (q, J=7.1 Hz, 2H), 1.26 (m, 12H), 1.2 (t, J=7.2 Hz, 3H).

Preparation 62-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]aceticacid

Charge a 2000 mL RBF with ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate(39.15 g, 132.8 mmol), 2,4-dichloro-6-(trifluoromethyl)pyrimidine (27.9g, 126 mmol), 1,4-dioxane (800 mL), aqueous Na₂CO₃ (2 M, 200 mL, 400mmol) and bis(triphenylphosphine)palladium(II) dichloride (2.8 g, 4.0mmol). Heat the mixture to 85° C. After 2 h, cool the mixture to RT.Divide the reaction mixture into two portions and carry forwardaccording to the following methods:

Method A:

To the first portion of the reaction mixture add[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (18.69 g, 59.75 mmol) and heat to 70°C. for 2.5 h. Cool to RT. Add aqueous NaOH (2 M, 166 mL, 332 mmol) andstir overnight at RT. Add EtOAc (500 mL) and stir the mixture for 30min. Acidify the mixture to pH=7 using aqueous HCl (5 M) and extractwith EtOAc (4×250 mL). Combine the organics, wash with saturated aqueousNaCl, dry over Na₂SO₄, filter and concentrate in-vacuo. Dissolve theresidue in DCM (125 mL) and add heptane (125 mL) dropwise. Stir themixture for 30 min. Collect the solid by filtration and wash with 1:1DCM/heptane (50 mL). Air-dry the solid.

Method B:

Concentrate the second portion of the reaction mixture in-vacuo toremove the 1,4-dioxane. Partition the mixture between EtOAc and water.Separate the aqueous layer and concentrate the organic layer to dryness.To the flask containing the residue add 1,4-dioxane (420 mL), aqueousNa₂CO₃ (2 M, 75 mL, 150 mmol) and[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (18.69 g, 59.75 mmol). Heat the mixtureto 70° C. and stir for 1.5 h. Cool the mixture to RT. Add aqueous NaOH(2 M, 125 mL, 250 mmol) and stir overnight. Add EtOAc (500 mL) and stirfor 30 min. Separate the aqueous phase and allow it to sit overnight.Acidify the aqueous mixture to pH=7 using aqueous HCl (5 M) and extractwith EtOAc (2×250 mL). Combine the organics, wash with saturated aqueousNaCl, dry over Na₂SO₄, filter and concentrate in-vacuo to give a solid.

Purification Method:

Combine the products from Methods A and B and dissolve in THE (485 mL).Add SiliaMetS® Thiol resin (32 g). Stir the mixture for 1 h and thenfilter. Concentrate the filtrate in-vacuo to give the title compound asa white powder (31.7 g, 72%). ES/MS (m/z): 342 (M+H); 340 (M−H).

Preparation 7 tert-Butyl4-[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]piperazine-1-carboxylate

Charge a 1 L RBF with2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]aceticacid (31.7 g, 91.0 mmol), tert-butyl piperazine-1-carboxylate (20.5 g,109 mmol) and DCM (320 mL). To the resulting solution add Et₃N (25.6 mL,182 mmol) and then dropwise2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxideanhydride solution (50 wt % in EtOAc, 68 mL, 114.2 mmol). Stir at RT for1 h. Wash the reaction with water (500 mL) and then saturated aqueousNaCl. Dry the organics over Na₂SO₄, filter, and concentrate under highvacuum to give the title compound (52.5 g, estimated 88 wt % pure basedon theoretical quantitative yield of product). ES/MS (m/z): 510 (M+H);508 (M−H).

Preparation 8 tert-Butyl4-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

To a microwave vial add 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.7g, 3.05 mmol) and 1,4-dioxane (15 mL). To the solution add tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate(1.55 g, 4.0 mmol), tetrakis(triphenylphosphine)palladium(0) (0.195 g,0.17 mmol) and aqueous Na₂CO₃ (2 M, 5.5 mL, 11 mmol). Seal the vial andheat in a microwave reactor at 85° C. for 1 h. In a second vial, carryout the same reaction at the same scale. Combine the reaction mixturesin a separatory funnel. Dilute the reaction mixture with saturatedaqueous sodium bicarbonate and extract twice with EtOAc. Combine theextracts and dry over Na₂SO₄, filter, and concentrate. Purify theresidue by silica gel chromatography using a gradient of 0 to 50%EtOAc/hexanes to give the title compound as a white solid (2.33 g, 88%).ES/MS (m/z): 432, 434 (M+H); 430, 432 (M−H).

Preparation 9 tert-Butyl4-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

In three separate microwave reaction vials, combine tert-butyl4-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate(1 g, 2.3 mmol) and 1,4-dioxane (16 mL). Add (R)-2-azetidinemethanolhydrochloride (0.52 g, 4.3 mmol) and DIPEA (1.6 mL, 9.3 mmol). Seal thevials and heat in a microwave reactor to 130° C. for 2.5 h. Combine theresulting reaction mixtures, then dilute the mixture with saturatedaqueous NaHCO₃ and extract twice with EtOAc. Combine the extracts anddry over Na₂SO₄, filter, and concentrate. Purify the residue by silicagel chromatography using a gradient of 10 to 80% EtOAc/hexanes to givethe title compound as a white foam (3.2 g, 95%). ES/MS (m/z): 483 (M+H).

Preparation 10 tert-Butyl3-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

To a microwave vial add 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.50g, 2.30 mmol) and 1,4-dioxane (22 mL) and water (2 mL). To the solutionadd tert-butyl3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1H-pyrazole-1-yl)azetidine-1-carboxylate(0.630 g, 1.71 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.20 g,0.26 mmol) and K₂CO₃ (500 mg, 3.67 mmol). Seal the vial and stir at RTovernight. Load the crude reaction on a silica cartridge, dry in vacuumoven and purify the residue by flash chromatography (gradient 0 to 45%EtOAc/hexane) to give the title compound as a light brown oil (0.375 g,49%). ES/MS (m/z): 348 (M+H-^(t)Bu); 402 (M−H).

Preparation 11 tert-Butyl3-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

To a vial combine tert-butyl3-[4-[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate(190 mg, 0.47 mmol) and THE (18 mL). Add (R)-2-azetidinemethanolhydrochloride (150 mg, 1.23 mmol) and DIPEA (1.0 mL, 5.7 mmol). Seal thevial and heat in the microwave reactor to 100° C. for 1 h 40 min. Aftercooling to RT, load the crude reaction on a silica cartridge, dry invacuum oven and purify the residue by silica gel chromatography using agradient of 0 to 80% (5% MeOH/EtOAc)/hexanes to give the title compoundas a colorless oil (153 mg, 72%). ES/MS (m/z): 455 (M+H).

Preparation 12 tert-Butyl3-[4-[2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

Prepare the title compound essentially as described in Preparation 11using (2S,3R)-2-methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid. ES/MS(m/z): 455 (M+H).

Preparation 13 tert-Butyl3-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate

Prepare the title compound essentially as described in Preparation 11using [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt. Purify the reaction by silica gelchromatography using 45% EtOAc/hexanes. ES/MS (m/z): 439 (M+H).

Preparation 142-[4-[5-Cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]aceticacid

Charge an RBF with ethyl2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl)acetate(2.95 g, 10.5 mmol),2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (2 g, 8.3 mmol),1,4-dioxane (52.4 mL) and aqueous Na₂CO₃ (2 M, 13.3 mL, 26.6 mmol).Sparge the mixture with nitrogen for 10 min. Addbis(triphenylphosphine)palladium(II) dichloride (191 mg, 0.27 mmol).Stir the mixture at RT for 16.5 h. Equip the flask with a refluxcondenser and add[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (2.73 g, 9.10 mmol). Heat the mixtureto 70° C. and stir for 3 h, then cool to RT. Add aqueous NaOH (2 M, 21mL, 42 mmol) and stir the mixture at RT for 15 min. Evaporate theorganic solvent in-vacuo. Dilute the residue with water (50 mL) anddecant the water away from the solid. Repeat this process two times withwater (50 mL). Collect the solid by vacuum filtration and air-dryovernight. Add 2-methyltetrahydrofuran (30.4 mL) and aqueous citric acid(6.5 wt %, 30.4 mL). Stir for 5 min and then separate the layers. Washthe organic layer with aqueous citric acid (6.5 wt %, 30.4 mL). Dry theorganic solution over MgSO₄, filter, and concentrate to give the titlecompound (3.14 g, estimated 97 wt % pure based on theoreticalquantitative yield of product). ES/MS (m/z): 366 (M+H); 364 (M−H).

Preparation 15 tert-Butyl4-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate

Method A:

To a microwave reactor vial add2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (497 mg, 2.063mmol), tert-butyl4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylate(867.2 mg, 2.3 mmol),1,1′-bis(diphenylphosphino)ferrocene-palladium(II)dichloride DCM complex(91.2 mg, 0.109 mmol), aqueous potassium carbonate (3 M solution, 2.1mL, 6.3 mmol), and 1,4-dioxane (10.5 mL). Sparge the mixture withnitrogen for 5 min, seal, and heat to 80° C. After 1 h, cool to RT.Filter the mixture over a Celite® plug, rinsing with EtOAc. Concentrateand purify the residue by silica gel chromatography using a gradient of15 to 40% EtOAc in hexanes, then briefly dry in-vacuo at 35° C. to givethe title compound (705 mg, 64%). ES/MS (m/z): 454, 456 (M−H).

Method B:

To an RBF add 2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile(1.07 g, 4.44 mmol) and 1,4-dioxane (25 mL). Add tert-butyl4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylate(2.08 g, 5.51 mmol), tetrakis(triphenylphosphine)palladium(0) (200 mg,0.17 mmol) and aqueous Na₂CO₃ (2 M, 8.5 mL, 17 mmol). Degas the reactionwith nitrogen and heat the reaction at 85° C. for 1.5 h. Decant theorganic phase from the reaction and split the crude reaction into twoequal amounts to use in Preparations 16 and 17. ES/MS (m/z): 454 (M−H).

Preparation 16 tert-Butyl4-[4-[5-cyano-6-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate

In a microwave vial add 12 of the crude reaction product fromPreparation 15, Method B, (R)-2-azetidinemethanol hydrochloride (0.5 g,4 mmol), DIPEA (1.6 mL, 9.2 mmol) and additional amount of 1,4-dioxane(16 mL). Heat the reaction at 120° C. for 2 h. Cool and concentrate thereaction to a crude product. Purify the residue by silica gelchromatography using a gradient of 50 to 80% EtOAc/hexanes, and purifyagain by silica gel chromatography using a gradient of 2 to 5% IPA/DCMto give the title compound as a tan solid (264 mg, 26%). ES/MS (m/z):507 (M+H).

Preparation 17 tert-Butyl4-[4-[5-cyano-6-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate

Prepare the title compound essentially as described in Preparation 16using the second ½ of the crude reaction product from Preparation 15 and(2S,3R)-2-methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid. ES/MS(m/z): 507 (M+H).

Preparation 182-Chloro-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrilehydrochloride

To a vial add tert-butyl4-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate(705 mg, 1.5 mmol), 4 M HCl in 1,4-dioxane (1.5 mL, 6.0 mmol), and DCM(1.5 mL). Stir the mixture at RT. After 1 h, concentrate the mixturein-vacuo and dry the residue under vacuum overnight to give the titlecompound (608 mg), which is used as-is without further purification inPreparations 19 and 20. ES/MS (m/z): 356, 358 (free base M+H).

Preparation 192-Chloro-6-[1-(1-methyl-4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

To a vial add2-chloro-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrilehydrochloride (303.3 mg, 0.74 mmol), 13.3 M formaldehyde in water (0.2mL, 3 mmol), sodium triacetoxyborohydride (378 mg, 1.73 mmol), and1,2-dichloroethane (6.0 mL). Stir the mixture at RT for 30 min. Addsaturated aqueous sodium bicarbonate and extract with DCM two times.Combine the extracts and concentrate in-vacuo to give the title compound(285 mg, 99% yield). ES/MS (m/z): 370, 372 (M+H).

Preparation 20 tert-Butyl4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate

Prepare the title compound from2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile and tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylateusing essentially the same procedure as Example 10 below. ES/MS (m/z):491 (M+H)

Preparation 21 tert-Butyl4-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate

Dissolve 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.258 g, 1.1 mmol)in 1,4-dioxane (6 mL) and add tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate(0.591 g, 1.52 mmol), tetrakis(triphenylphosphine)palladium(0) (68 mg,0.059 mmol) and 2M aqueous sodium carbonate (2.2 mL, 4.4 mmol). Heat themixture to 85° C. in a microwave reactor for 1 h. Cool to RT and add[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (0.414 g, 1.36 mmol) and DIPEA (0.6 mL,3.4 mmol). Heat the mixture for 2 h at 110° C. Dilute the mixture withsaturated aqueous sodium bicarbonate and extract twice with EtOAc.Combine the extracts, dry over sodium sulfate, then filter andevaporate. Purify the residue by silica gel chromatography using agradient of 0 to 50% EtOAc/hexanes to give the title compound (215 mg,41%) as a white solid. ES/MS (m/z): 467 (M+H).

Preparation 22 tert-Butyl3-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]azetidine-1-carboxylate

To a vial add 3-cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (0.360 g,1.49 mmol), 1,4-dioxane (7.5 ml), tert-butyl3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate(0.727 g, 1.98 mmol), tetrakis(triphenylphosphine)palladium(0) (95 mg,0.08 mmol), and 2M aqueous sodium carbonate (2.8 mL, 5.6 mmol, 2 mol/L).Seal the vessel and heat in a microwave reactor at 85° C. for 1 h.Dilute with saturated sodium bicarbonate and extract twice with EtOAc.Combine the extracts, dry over sodium sulfate, filter and evaporate.Purify the residue by silica gel chromatography using a gradient of 0 to90% EtOAc/hexanes to give the title compound (0.609 g, 95%). ES/MS(m/z): 426, 428 (M−H).

Preparation 232-Chloro-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

Prepare the title compound from1-tetrahydropyran-4-yl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazoleusing essentially the same procedure as Preparation 22. ES/MS (m/z):357, 359 (M+H).

Preparation 24 tert-Butyl3-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]azetidine-1-carboxylate

Dissolve tert-butyl3-[4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]azetidine-1-carboxylate(603 mg, 1.410 mmol) in THE (18 mL). Add[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (519 mg, 1.711 mmol) and DIPEA (0.75mL, 4.3 mmol), then heat the mixture to 130° C. for 2.5 h. Evaporate thesolvent and purify the residue by silica gel chromatography using agradient from 0 to 50% EtOAc/hexanes to give the title compound (498 mg,76%) as a colorless oil. ES/MS (m/z): 463 (M+H), 461 (M−H).

Preparation 25 tert-ButylN-[2-[4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-1-piperidyl]-2-oxo-ethyl]carbamate

Dissolve2-[(2S)-2-methylazetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(0.129 g, 0.330 mmol) in DCM (1 mL). Add2-(tert-butoxycarbonylamino)acetic acid (69 mg, 0.39 mmol), HATU (0.175g, 0.460 mmol) and DIPEA (0.18 mL, 1.0 mmol). Stir the mixture at RT for3 h, then dilute with saturated aqueous sodium bicarbonate and extracttwice with DCM. Combine the organics, dry over sodium sulfate, thenfilter and evaporate. Purify the residue by reverse-phase chromatographyon silica-bound C18 (solvent A=10 mM aqueous ammonium bicarbonate,solvent B=ACN; gradient 10 to 91% B) to give the title compound (129 mg,71%). ES/MS (m/z): 548 (M+H).

Preparation 261-(2,2-Dimethyl-[1,3]dioxolan-4-ylmethyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole

Suspend 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole(0.500 g, 2.50 mmol) in DMF (10 mL), then add4-(chloromethyl)-2,2-dimethyl-1,3-dioxolane (0.73 mL, 5.0 mmol) andcesium carbonate (1.64 g, 5.03 mmol). Heat the mixture to 75° C.overnight. Dilute the reaction with saturated aqueous sodium bicarbonateand extract with EtOAc. Combine the organics and wash four times withsaturated aqueous NaCl, then dry over sodium sulfate, filter andevaporate to give the title compound (780 mg), which is carried forwardwithout further purification. ES/MS (m/z): 309 (M+H).

Preparation 272-Chloro-6-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve 3-cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (0.352 g, 1.46mmol) in 1,4-dioxane (7.3 mL). Add1-(2,2-dimethyl-[1,3]dioxolan-4-ylmethyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-pyrazole(0.602 g, 1.89 mmol), tetrakis(triphenylphosphine)palladium(0) (0.715 g,0.619 mmol) and aqueous 2 M sodium carbonate (2.8 mL, 5.6 mmol). Purgethe mixture with nitrogen for 15 min, then heat the mixture to 85° C.for 1.5 h. Dilute the reaction with saturated aqueous sodium bicarbonateand extract twice with EtOAc. Combine the organics and dry over sodiumsulfate, then filter and evaporate. Purify the residue by silica gelchromatography using a gradient of 0 to 40% EtOAc/hexanes to give thetitle compound (207 mg, 36%) as a colorless oil. ES/MS (m/z): 387, 389(M+H).

Preparation 28 tert-Butyl4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazole-1-carboxylate

In a vial combine 3-cyano-2,6-dichloro-4-(trifluoromethyl)pyridine (195mg, 0.785 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate(250 mg, 0.85 mmol), 2 M aqueous sodium carbonate (2.5 mL, 5.0 mmol),and 1,4-dioxane (4 mL). Degas the reaction at RT by bubbling nitrogenthrough the reaction with stirring for 5 min. Add[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (185 mg,0.240 mmol) and heat the reaction to 100° C. for 2 h. Add water andextract with EtOAc. Combine the extracts and concentrate in-vacuo.Purify the residue by silica gel chromatography using a gradient of 25to 50% EtOAc/hexanes to give the title compound (40 mg, 14%) as anorange solid. ES/MS (m/z): 271, 273 (M−H—BOC).

Preparation 29 tert-ButylN-[2-[[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]amino]ethyl]carbamate

To a vial add2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]aceticacid (200 mg, 0.586 mmol), DMF (2 mL), DIPEA (0.31 mL, 1.76 mmol), HATU(0.267 g, 0.703 mmol), and N—BOC-ethylenediamine (0.102 mL, 0.644 mmol).Stir for 8 h at RT, then purify by preparative HPLC [parameters:solvents—aqueous 10 mM ammonium bicarbonate pH 10/5% MeOH (Solvent A)and ACN (Solvent B); precolumn—Waters BEH HILIC 100×30 mm 5 μm, 110 Åwith a 15×30 mm BEH HILIC guard column; column: Phenomenex® Kinetex® EVOC18, 100×30 mm, 5 μm, 100 Å with a 15×30 mm EVO guard column usinginline heater at 50° C.; gradient 33 to 67% B] to give the titlecompound (179 mg, 63%). ES/MS (m/z): 484 (M+H).

Prepare the compounds shown in Table 1 using essentially the sameprocedure as Preparation 29 and the appropriate commercially availableamine.

TABLE 1 Prepa- ES/MS ration (m/z): Number Name Structure (M + H) 30tert-butyl (2S)-2- methyl-4-[2-[4-[2-[(2S)- 2-methylazetidin-1-yl]-6-(trifluoromethyl) pyrimidin-4-yl]pyrazol- 1-yl]acetyl]piperazine-1-carboxylate

524 31 tert-butyl (3S)-3- methyl-4-[2-[4-[2-[(2S)-2-methylazetidin-1-yl]- 6-(trifluoromethyl) pyrimidin-4-yl]pyrazol-1-yl]acetyl]piperazine- 1-carboxylate

524 32 tert-butyl (2R,5S)-2,5- dimethyl-4-[2-[4-[2-[(2S)-2-methylazetidin- 1-yl]-6- (trifluoromethyl) pyrimidin-4-yl]pyrazol-1-yl] acetyl]piperazine-1- carboxylate

538 33 tert-butyl 2,2-dimethyl- 4-[2-[4-[2-[(2S)-2-methylazetidin-1-yl]- 6-(trifluoromethyl) pyrimidin-4-yl]pyrazol-1-yl]acetyl] piperazine-1- carboxylate

538

Preparation 34 tert-Butyl(3R)-3-[4-[2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylate

In a reaction vessel, combine 2,4-dichloro-6-(trifluoromethyl)pyrimidine(600 mg, 2.71 mmol) with (R)-tert-butyl3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate(1 g, 2.67 mmol), 2 M aqueous sodium carbonate (3 mL, 6 mmol) andbis(triphenylphosphine)palladium(II) dichloride (50 mg, 0.068 mmol) in1,4-dioxane (6 mL). Degas with nitrogen and heat the mixture to 80° C.for 2 h. Dilute with EtOAc (75 mL), wash with water and saturatedaqueous NaCl. Dry the organic layer over sodium sulfate, filter andconcentrate. Combine the residue with (2S,3R)-2-methylazetidin-3-ol[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonic acid (420 mg,1.32 mmol) in 1,4-dioxane (4 mL) and DIPEA (0.7 mL, 4 mmol) in a vial.Seal the vessel and heat to 120° C. in a microwave reactor for 1 h. Loadthe reaction mixture directly onto silica gel and purify by silica gelchromatography using a gradient from 0 to 80% EtOAc/hexanes to give thetitle compound (570 mg, 92%) as an off-white foam. ES/MS (m/z): 469[M+H].

Preparation 35 tert-Butyl(3S)-3-[4-[2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylate

Prepare the title compound essentially as described in Preparation 34using (S)-tert-butyl3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)pyrrolidine-1-carboxylate.ES/MS (m/z): 469 [M+H].

Preparation 36 tert-Butyl3-[(1R)-4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]pyrrolidine-1-carboxylate

Prepare the title compound essentially as described in Preparation 34using (R)-2-azetidinemethanol hydrochloride. ES/MS (m/z): 469 [M+H].

Preparation 37 tert-butyl4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-3-hydroxy-piperidine-1-carboxylate

To a vial add4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (560 mg, 2.9mmol), tert-butyl 7-oxa-4-azabicyclo[4.1.0]heptane-4-carboxylate (580mg, 2.9 mmol), DMF (10 mL), and cesium carbonate (1.7 g, 5.2 mmol). Heatthe mixture to 80° C. for 6 h. Dilute the reaction with water andextract with EtOAc. Combine the extracts and dry over sodium sulfate,filter and concentrate to give crude tert-butyl3-hydroxy-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]piperidine-1-carboxylate(650 mg). Place this material in a RBF and add2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (300 mg, 1.2mmol), 1,4-dioxane (5 mL), and aqueous 2 M sodium carbonate (1.3 mL, 2.6mmol). Degas the reaction at RT by bubbling nitrogen through thereaction with stirring for 5 min. Addtetrakis(triphenylphosphine)palladium(0) (80 mg, 0.06 mmol) and degasfor an additional 3 min. Heat the reaction to 80° C. for 4 h, then coolto RT. Add DIPEA (0.6 mL, 3 mmol) and[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (410 mg, 1.4 mmol). Stir for 30 min atRT and then heat to 80° C. for 1 h. Dilute the mixture with water andextract with EtOAc. Combine the extracts and dry over sodium sulfate,filter and concentrate. Purify the residue by reverse phasechromatography (C18, gradient 10 to 100% ACN/aqueous 10 mM ammoniumcarbonate+5% methanol) to give the title compound (95 mg, 15%). ES/MS(m/z): 507 (M+H).

Example 12-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone

Charge a 2 L RBF with tert-butyl4-[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]piperazine-1-carboxylate(47.0 g, 89.5 mmol) and add DCM (470 mL). Add TFA (60 mL, 778 mmol)dropwise to the mixture and stir at RT for 19 h. Add more TFA (8 mL, 110mmol) and continue stirring for 18 h. Slowly add the reaction mixture toa flask containing chilled aqueous NH₄OH (35 wt %, 150 mL, 1300 mmol).Separate the layers. Wash the organic layer with saturated aqueous NaCl,dry over Na₂SO₄, filter, and concentrate. Dissolve the residue in EtOAc(400 mL) and concentrate in-vacuo. Dissolve the residue in IPA (400 mL)and concentrate in vacuo to give the title compound as an off-white foam(32 g, 84%). ES/MS (m/z): 410 (M+H).

Example 1a2-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanonesesquisuccinate

Add2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-piperazin-1-yl-ethanone(959 g, 2.34 mol) and then IPA (11.5 L) to a reactor. Add succinic acid(550 g, 4.69 mol) and heat the mixture to 70-80° C. to give a solution.Stir the mixture between 70 and 80° C. for 2 h, then cool to 25° C. over6 h. Filter the mixture and rinse with IPA (1 L). Dry the resultingsolids at 40-50° C. for 6 h to provide the title compound (1070 g, 78%)as a white solid. ¹H-NMR (400 MHz, DMSO-d₆) δ 8.41 (s, 1H), 8.18 (s,1H), 7.33 (s, 1H), 5.24 (s, 2H), 4.53-4.43 (m, 1H), 4.06-3.93 (m, 2H),3.58-3.47 (m, 4H), 2.95-2.82 (m, 4H), 2.47-2.40 (m, 1H), 2.37 (s,6H-succinate methylene groups, 1.5 equiv), 2.02-1.92 (m, 1H), 1.50 (d,J=4 Hz, 3H). High-resolution ES-MS (m/z): theoretical 410.1911 (freebase M+H), observed 410.1916.

XRPD peaks of Example 1a are listed in Table 2.

TABLE 2 X-ray powder diffraction peaks of Example 1a Relative IntensityAngle (% of most Peak (°2-Theta) intense peak) 1 12.1 27.5% 2 14.1 22.7%3 16.3 33.3% 4 17.7 30.5% 5 18.9 100.0% 6 20.0 22.1% 7 20.5 53.8% 8 21.344.9% 9 22.0 36.6% 10 24.3 23.2%

Example 2[(2R)-1-[4-[1-(4-Piperidyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

Dissolve tert-butyl4-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate(2.55 g, 5.28 mmol) in DCM (50 mL) and add TFA (10 mL, 132 mmol). Stirthe mixture at RT for 30 min then apply it directly to SCX resin. Washthe resin with MeOH and then with ammonia in MeOH solution (7 M).Combine the fractions containing the desired product and concentrate invacuo. Purify the residue by silica gel chromatography using a gradientfrom 0 to 9% (7 M ammonia/MeOH)/DCM to give the title compound as awhite powder (2.02 g, 94%). ES/MS (m/z): 383 (M+H); 381 (M−H).

Example 3[(2R)-1-[4-[1-(Azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

Dissolve tert-butyl3-[4-[2-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate(153 mg, 0.336 mmol) in DCM (15 mL) and add TFA (5 mL, 66 mmol). Stirthe mixture at RT for 1 h then concentrate under reduced pressure at RT.Co-evaporate the residue with DCM and dry under vacuum. Purify theresidue by preparative HPLC (parameters: Solvent A=aqueous 10 mMammonium bicarbonate pH 10/5% MeOH, Solvent B=ACN; column—Phenomenex®Kinetex® EVO C18, 100×30 mm, 5 um, 100 Å with 15×30 mm EVO guard column)to give the title compound (67 mg, 40%) as a white powder. ES/MS (m/z):355 (M+H); 353 (M−H).

Example 4(2S,3R)-1-[4-[1-(Azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol

Dissolve tert-butyl3-[4-[2-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate(153 mg, 0.336 mmol) in DCM (15 mL) and add TFA (5 mL, 66 mmol). Stirthe mixture at RT for 1 h then concentrate under reduced pressure at RT.Co-evaporate the residue with DCM and dry under vacuum. Purify theresidue by preparative HPLC (parameters: Solvent A=aqueous 10 mM NH₄HCO₃with 5% MeOH pH 10, Solvent B=CAN; column—Phenomenex® Kinetex® EVO C18,100×30 mm, 5 um, 100 Å with a 15×30 mm EVO guard column to give thetitle compound (79 mg, 57%) as a white powder. ES/MS (m/z): 355 (M+H);353 (M−H).

Example 52-[4-[5-Cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-N-(2-hydroxyethyl)acetamide

Dissolve2-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]aceticacid (351 mg, 0.96 mmol) in DMF (2 mL) and add HATU (482 mg, 1.24 mmol),ethanolamine (0.1 mL, 2 mmol) and DIPEA (0.5 mL, 3 mmol). Stir themixture at RT for 2 d. Purify the reaction mixture directly byreversed-phase chromatography (C18-bonded silica) using a gradient from5 to 95% MeCN/aqueous (NH₄)₂CO₃ (10 mM) to give the title compound (110mg, 28%). ES/MS (m/z): 409 (M+H); 407 (M−H).

Example 62-[(2R)-2-(Hydroxymethyl)azetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve tert-butyl4-[4-[5-cyano-6-[(2R)-2-(hydroxymethyl)azetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate(264 mg, 0.52 mmol) in DCM (2 mL). Add TFA (2 mL) in one portion to thereaction and stir for 1 h. Concentrate the reaction, dilute with DCM andconcentrate the reaction mixture. Dilute the reaction material with MeOH(1 mL) and add saturated aqueous NaHCO₃ to bring the pH-8. Purify thereaction mixture using reverse phase chromatography (40 g, C18, gradient20-100% ACN/aqueous 10 mM (NH₄)₂CO₃ with 5% MeOH) to give the titlecompound (135.5 mg, 64%). ES/MS (m/z): 407 (M+H).

Example 72-[(2S,3R)-3-Hydroxy-2-methyl-azetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve tert-butyl4-[4-[5-cyano-6-[(2S,3R)-3-hydroxy-2-methyl-azetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate(220 mg, 0.43 mmol) in DCM (2 mL). Add TFA (2 mL) in one portion to thereaction and stir for 1 h. Concentrate the reaction, dilute with DCM andconcentrate the reaction mixture. Dilute the reaction material with MeOH(1 mL) and add saturated aqueous NaHCO₃ to bring the pH-2. Purify thereaction mixture using reverse phase chromatography (40 g, C18, gradient20-100% ACN/aqueous 10 mM (NH₄)₂CO₃ with 5% MeOH) to give the titlecompound (127 mg, 72%). ES/MS (m/z): 407 (M+H).

Example 82-[(2S)-2-Methylazetidin-1-yl]-6-[1-(1-methyl-4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

To a vial add2-chloro-6-[1-(1-methyl-4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(261 mg, 0.68 mmol),[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (338 mg, 1.1 mmol), DIPEA (0.4 mL, 2mmol), and THF (3.7 mL). Seal the vial and heat to 130° C. for 2 h.Partition the reaction mixture between DCM and aqueous 1 N HCl.Neutralize the aqueous phase with 1 N NaOH, then extract three timeswith DCM. Combine the organic extracts. Dry over sodium sulfate, filter,and concentrate in-vacuo. Dry the material at 50° C. in a vacuum ovenfor 1 h. Purify the residue by silica gel chromatography using agradient of 5 to 10% MeOH/DCM, combine fractions containing the titlecompound, concentrate and dry in a vacuum oven at 50° C. overnight.Purify the residue by preparative HPLC (parameters: Solvent A=10 mMaqueous ammonium bicarbonate with 5% MeOH, Solvent B=ACN;column—Xbridge™ 30 mm×75 mm 5 μm, 45 mL/min; gradient—5 to 100% B) togive the title compound (73 mg, 26%). ES/MS (m/z): 405 (M+H).

Example 96-[1-[1-(2-Hydroxyethyl)-4-piperidyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

To a 20 mL vial add2-chloro-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrilehydrochloride (304.3 mg, 0.63 mmol),2-[tert-butyl(dimethyl)silyl]oxyacetaldehyde (219 mg, 1.19 mmol), DIPEA(0.52 mL, 3.1 mmol), sodium triacetoxyborohydride (277.2 mg, 1.3 mmol)and DCM (2.0 mL). Stir the reaction mixture at RT for 2 h. Add2-[tert-butyl(dimethyl)silyl]oxyacetaldehyde (219 mg, 1.2 mmol) and stirovernight at RT. Add a saturated solution of aqueous sodium bicarbonateand extract two times with DCM. Combine the organic phases, wash withsaturated aqueous NaCl, dry over sodium sulfate, filter, and concentratein-vacuo. Purify the residue by silica gel chromatography using agradient of 40 to 70% EtOAc in hexanes to give6-[1-[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]pyrazol-4-yl]-2-chloro-4-(trifluoromethyl)pyridine-3-carbonitrile(220 mg, 58%).

To a vial add6-[1-[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]pyrazol-4-yl]-2-chloro-4-(trifluoromethyl)pyridine-3-carbonitrile(175 mg, 0.289 mmol),[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (136 mg, 0.45 mmol), DIPEA (0.15 mL,0.85 mmol), and THE (1.5 mL, 18 mmol). Seal the vessel and heat to 130°C. for 60 min. Partition the reaction mixture between EtOAc and 1 N HCl,remove the organic layer, and extract the aqueous layer three times withEtOAc. Combine the organics, wash with saturated aqueous NaCl, dry oversodium sulfate, filter, and concentrate in-vacuo. Dissolve the residuein THE (1.3 mL) and cool the mixture to 0° C., then add 1 Mtetrabutylammonium fluoride in THE (0.37 mL, 0.37 mmol) drop-wise. Allowthe reaction to warm to RT and stir overnight. Concentrate the reactionmixture in-vacuo. Purify the residue by silica gel chromatography using100% hexanes, then using a gradient of 1 to 10% (0.7 N ammonia inMeOH)/DCM to give the title compound (93 mg, 73%). ES/MS (m/z): 435(M+H).

Example 106-[1-[2-(Dimethylamino)ethyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

To a reaction vessel add2,6-dichloro-4-(trifluoromethyl)pyridine-3-carbonitrile (250 mg, 1.04mmol),N,N-dimethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanamine(271 mg, 1.07 mmol), 1,4-dioxane (10 mL), aqueous potassium carbonate (3M, 1.04 mL, 3.12 mmol), and[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)) (40 mg,0.052 mmol). Seal the vessel and heat to 80° C. for 2 h, then cool themixture to RT. Filter through a 3 g cartridge of Celite® using EtOAc toelute. Concentrate the filtrate by evaporation. To the residue add[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (378 mg, 1.24 mmol), DIPEA (0.9 mL, 5.2mmol), and DMSO (10 mL). Heat the mixture to 120° C. for 6 h. Dilute thereaction mixture with water and load onto a Strata-XL® cartridge (8 g;previously washed with MeOH, dried, and then washed with water). Washthe cartridge with water followed by 1:1 MeOH/water and discard theelutents. Elute the product with MeOH followed by DCM and finally a 1:1mixture of DCM/MeOH, pooling the eluted fractions. Concentrate thepooled fractions containing the title compound. Purify the residue bypreparative HPLC (parameters: Solvent A=10 mM aqueous ammoniumbicarbonate with 5% MeOH pH 10, Solvent B=ACN; precolumn—Waters BEHHILIC 100×30 mm 5 μm, 110 Å with a 15×30 mm BEH HILIC guard column;column—Phenomenex® Kinetex® EVO C18, 100×30 mm, 5 μm, 100 Å with a 15×30mm EVO guard column using inline heater at 50° C.; gradient 33 to 100%B) to give the title compound (53 mg, 13%). ES/MS (m/z): 379 (M+H).

Example 116-[1-(2-Hydroxyethyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Prepare the title compound using essentially the method described inExample 10 with2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)ethanol.ES/MS (m/z): 352 (M+H).

Example 122-[(2S)-2-Methylazetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve tert-butyl4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]piperidine-1-carboxylate(1.99 g, 4.06 mmol) in DCM (50 mL) and add TFA (10 mL, 132.3 mmol)slowly. Stir at RT for 30 min then load the reaction mixture onto four10 g SCX cartridges. Wash the cartidges with MeOH and then with 7Nammonia in MeOH. Concentrate the basic washes in-vacuo. Purify theresidue by reverse-phase chromatography on silica-bound C18 (Solvent A:10 mM ammonium bicarbonate with 5% MeOH; Solvent B: ACN; gradient:10-71% solvent B) to give the title compound (709 mg, 45%) as a whitesolid. ES/MS (m/z): 391 (M+H).

Example 132-[(2S)-2-Methylazetidin-1-yl]-4-[1-(4-piperidyl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine

Dissolve tert-butyl4-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]piperidine-1-carboxylate(0.213 g, 0.457 mmol) in DCM (5 mL) and add TFA (1 mL). Stir the mixturefor 25 min then load the reaction mixture directly onto a 10 g SCXcartridge. Wash the cartridge with MeOH and then elute with 7 N ammoniain MeOH. Concentrate the basic wash to give the title compound (129 mg,77%). ES/MS (m/z): 367 (M+H).

Prepare the examples in Table 3 using essentially the method describedin Example 13 and the appropriate protected amine.

TABLE 3 ES/MS Example (m/z): Number Name Structure (M + H) 14(2S,3R)-2-Methyl-1-[4-[1-[(3R)- pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2- yl]azetidin-3-ol

369 15 (2S,3R)-2-Methyl-1-[4-[1-[(3S)- pyrrolidin-3-yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2- yl]azetidin-3-ol

369 16 [(2R)-1-[4-[1-[(3R)-Pyrrolidin-3- yl]pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2- yl]azetidin-2-yl]methanol

369

Example 176-[1-(Azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Prepare the title compound essentially using the same procedure asExample 12 starting from tert-butyl3-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]azetidine-1-carboxylate.Purify by preparative HPLC (parameters: solvent A—10 mM aqueous ammoniumbicarbonate with 5% MeOH, solvent B—ACN; gradient 35 to 59% B;column—Phenomenex® Kinetex® EVO C18, 100×30 mm, 5 μm) to give the titlecompound. ES/MS (m/z): 363 (M+H).

Example 186-[1-[1-(2-Aminoacetyl)-4-piperidyl]pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Prepare the title compound using essentially the same procedure asExample 12 starting from tert-butylN-[2-[4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-1-piperidyl]-2-oxo-ethyl]carbamate.Purify by preparative HPLC (parameters: solvent A—10 mM ammoniumbicarbonate with 5% MeOH; solvent B—MeOH; gradient—20 to 50% B, 55mL/min; column—Phenomenex® Kinetex® EVO C18 30 mm×250 mm, 5 m). ES/MS(m/z): 448 (M+H).

Example 196-[1-(1-Acetyl-4-piperidyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve2-[(2S)-2-methylazetidin-1-yl]-6-[1-(4-piperidyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(Example 12) (0.111 g, 0.284 mmol) in DCM (1 mL) and add pyridine (0.026mL, 0.32 mmol). Cool the mixture to 0° C. and add acetyl chloride (0.024mL, 0.34 mmol). Warm the mixture to RT and stir for 30 min. Cool themixture to 0° C. and add pyridine (0.026 mL, 0.32 mmol) and acetylchloride (0.024 mL, 0.34 mmol). Warm to RT and stir for an additional 30min. Dilute the reaction with saturated aqueous sodium bicarbonate andextract twice with DCM. Combine the organics, dry over sodium sulfate,filter and evaporate. Purify the residue by preparative HPLC(parameters: solvent A—10 mM ammonium bicarbonate with 5% MeOH; solventB—MeOH; gradient 25-55%, 55 mL/min; column—Phenomenex® Kinetex® EVO C1830 mm×250 mm, 5 m) to give the title compound (67 mg, 55%). ES/MS (m/z):433 (M+H).

Examples 20 to 23

Prepare the examples in Table 4 using essentially the procedure fromExample 5 and the appropriate carboxylic acid and commercially availableamine.

TABLE 4 ES/MS Example (m/z): Number Name Structure (M + H) 202-[(2S)-2-Methylazetidin-1-yl]-6-[1- [2-(4-methylpiperazin-1-yl)-2-oxo-ethyl]pyrazol-4-yl]-4- (trifluoromethyl)pyridine-3- carbonitrile

448 21 2-[4-[5-Cyano-6-[(2S)-2- methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-N,N-bis(2-hydroxyethyl)acetamide

453 22 6-[1-[2-[(3R,4R)-3,4- Dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2- methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3- carbonitrile

451 23 6-[1-[2-[(3S,4S)-3,4- Dihydroxypyrrolidin-1-yl]-2-oxo-ethyl]pyrazol-4-yl]-2-[(2S)-2- methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3- carbonitrile

451

Example 246-[1-(2,3-Dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve2-chloro-6-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(0.204 g, 0.527 mmol) in THE (6 mL). Add aqueous hydrochloric acid (2 M,3 mL, 6 mmol) to the mixture and stir at RT for 1.5 h. Dilute themixture with saturated aqueous sodium bicarbonate and extract twice withEtOAc. Combine the organics and dry over sodium sulfate, then filter andevaporate to give crude2-chloro-6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(183 mg). Dissolve this crude material in its entirety in DMF (2 mL),then add [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (222 mg, 0.73 mmol) and DIPEA (0.37 mL,2.1 mmol). Heat the mixture in a microwave reactor at 130° C. for 2.5 h.Purify the reaction mixture by preparative HPLC (parameters: solventA—10 mM ammonium bicarbonate with 5% MeOH; solvent B—ACN; gradient 20 to50%, 60 mL/min; column—Phenomenex® Kinetex® EVO C18 30 mm×250 mm, 5 m)to give the title compound (102 mg, 51%). ES/MS (m/z): 382 (M+H).

Examples 24a and 24b Example 24a:6-[1-(2,3-Dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile—Isomer1 Example 24b:6-[1-(2,3-Dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile—Isomer2

Separate the isomers of6-[1-(2,3-dihydroxypropyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile(86 mg) using chiral SFC (parameters: column—Phenomenex® Lux®Cellulose-2, 21×250 mm; column temperature—40° C.; solvent—15% EtOH/CO₂,80 mL/min) to provide the title compounds [Isomer 1, first-elutingisomer: 35 mg, ES/MS (m/z): 382 (M+H); Isomer 2, second-eluting isomer:39 mg, ES/MS (m/z): 382 (M+H)]. Analytical chiral SFC (parameters:column—Phenomenex® Lux® Cellulose-2, 4.6×150 mm; solvent—15% EtOH/CO₂, 5mL/min): Isomer 1—retention time 3.10 min, 95.6% ee; Isomer 2—retentiontime 3.52 min—94.4% ee.

Example 252-[(2S)-2-Methylazetidin-1-yl]-6-(1H-pyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

In a vial combine tert-butyl4-[6-chloro-5-cyano-4-(trifluoromethyl)-2-pyridyl]pyrazole-1-carboxylate(75 mg, 0.20 mmol), 1,4-dioxane (1.5 mL), and EtOH (1 mL). Add DIPEA(130 mg, 1.0 mmol) and[(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (117 mg, 0.39 mmol). Heat the reactionin a microwave reactor at 200° C. for 2 h. Concentrate the reactionin-vacuo. Purify the residue by reverse-phase chromatography onsilica-bound C18 using a gradient of 10 to 100% ACN/water with 0.1%formic acid to give the title compound (35 mg, 41%) as a pale yellowsolid. ES/MS (m/z): 308 (M+H), 306 (M−H).

Example 262-[(2S)-2-Methylazetidin-1-yl]-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile

In a vial combine2-chloro-6-(1-tetrahydropyran-4-ylpyrazol-4-yl)-4-(trifluoromethyl)pyridine-3-carbonitrile(105 mg, 0.29 mmol), 1,4-dioxane (1 mL), EtOH (0.5 mL), DIPEA (0.2 mL, 1mmol), and [(1R,4S)-7,7-dimethyl-2-oxo-norbornan-1-yl]methanesulfonate(2S)-2-methylazetidin-1-ium salt (100 mg, 0.33 mmol). Heat the reactionin a microwave reactor at 150° C. for 2 h. Concentrate the reactionin-vacuo. Purify the residue by silica gel chromatography using agradient of 40 to 50% EtOAc/hexanes to give the title compound (88 mg,76%) as a light yellow solid. ES/MS (m/z): 392 (M+H).

Example 276-[1-(3-Hydroxy-4-piperidyl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile

Dissolve tert-butyl4-[4-[5-cyano-6-[(2S)-2-methylazetidin-1-yl]-4-(trifluoromethyl)-2-pyridyl]pyrazol-1-yl]-3-hydroxy-piperidine-1-carboxylate(85 mg, 0.17 mmol) in DCM (1 mL), then add TFA (0.5 mL) and stir for 5min. Concentrate the reaction in-vacuo. Purify the residue by reversephase chromatography (C18, gradient 20 to 100% ACN/aqueous 10 mMammonium carbonate+5% methanol) to give the title compound (27 mg, 40%)as a white solid. ES/MS (m/z): 407 (M+H).

Example 28N-(2-Aminoethyl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetamide

Dissolve tert-butylN-[2-[[2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]acetyl]amino]ethyl]carbamate(340 mg) in neat TFA (5 mL) and stir at ambient temperature. After 2min, quench with aqueous NaOH until basic. Extract with DCM and EtOAc.Combine the extracts, dry over sodium sulfate, filter, and concentrate.Purify the residue by preparative HPLC [parameters: solvents—10 mMaqueous ammonium bicarbonate pH 10/500 MeOH (Solvent A) and ACN (SolventB); precolumn—Waters BEH HILIC 100×30 mm 5 μm, 110 Å with a 15×30 mm BEHHILIC guard column; column—Phenomenex® Kinetex® EVO C18, 100×30 mm, 5μm, 100 Å with a 15×30 mm EVO guard column using an in-line heater at50° C.; gradient 14 to 48% B] to give the title compound (45 mg, 170%).ES/MS (m/z): 384 (M+H).

Prepare the compounds shown in Table 5 using essentially the sameprocedure as Example 28 and the appropriate protected amine.

TABLE 5 ES/MS Example (m/z): number Name Structure (M + H) 292-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(3S)-3-methylpiperazin-1- yl]ethanone

424 30 2-[4-[2-[(2S)-2-Methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]-1-[(2S)-2-methylpiperazin-1- yl]ethanone

424 31 1-(3,3-Dimethylpiperazin-1-yl)-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6- (trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]ethanone

438 32 1-[(2S,5R)-2,5-Dimethylpiperazin-1-yl]-2-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol- 1-yl]ethanone

438

Example 334-[1-(Azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidine

Dissolve tert-butyl3-[4-[2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl]pyrazol-1-yl]azetidine-1-carboxylate(530 mg, 1.21 mmol) in DCM (50 mL). Add TFA (6 mL) and stir the mixtureat RT for 1 h. Concentrate the mixture in-vacuo to give the crude titlecompound. Purify half of this material by preparative HPLC [parameters:solvents—aqueous 10 mM ammonium bicarbonate pH 10/5% MeOH (Solvent A)and ACN (Solvent B); precolumn—Waters BEH HILIC 100×30 mm 5 μm, 110 Åwith a 15×30 mm BEH HILIC guard column; column—Phenomenex® Kinetex® EVOC18, 100×30 mm, 5 μm, 100 Å with a 15×30 mm EVO guard column usingin-line heater at 50° C.; gradient 23 to 58% B] to give the titlecompound (162 mg, 80% yield from half of the starting material) as awhite solid. ES/MS (m/z): 339 (M+H).

Example 342-[(2S)-2-Methylazetidin-1-yl]-4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidine

Dissolve half of the crude4-[1-(azetidin-3-yl)pyrazol-4-yl]-2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidineprepared in Example 33 (0.6 mmol) in MeOH (6 mL). Add acetic acid (0.1mL) and 3.45 M aqueous formaldehyde (1 mL) and stir the mixture at RTfor 30 min. Add sodium triacetoxyborohydride (300 mg, 1.4 mmol) and stirfor the mixture 2 h. Concentrate the mixture in-vacuo and purify theresidue by preparative HPLC [parameters: solvents—aqueous 10 mM ammoniumbicarbonate pH 10/5% MeOH (Solvent A) and ACN (Solvent B);precolumn—Waters BEH HILIC 100×30 mm 5 μm, 110 Å with a 15×30 mm BEHHILIC guard column; column—Phenomenex® Kinetex® EVO C18, 100×30 mm, 5μm, 100 with a 15×30 mm EVO guard column using inline heater at 50° C.;gradient 33-67% B] to give the title compound (128 mg, 61% yield fromhalf of the starting material in Example 33) as a white solid. ES/MS(m/z): 353 (M+H).

Example 35(2S,3R)-2-Methyl-1-[4-[1-(1-methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-3-ol

Prepare the title compound using essentially the same procedure asExample 34 beginning with(2S,3R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]-2-methyl-azetidin-3-ol(Example 4). ES/MS (m/z): 369 (M+H), 367 (M−H).

Example 36[(2R)-1-[4-[1-(1-Methylazetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol

Prepare the title compound using essentially the same procedure asExample 34 beginning with[(2R)-1-[4-[1-(azetidin-3-yl)pyrazol-4-yl]-6-(trifluoromethyl)pyrimidin-2-yl]azetidin-2-yl]methanol(Example 3). ES/MS (m/z): 369 (M+H), 367 (M−H).

Assays KHK Enzyme Activity Assay for Human KHK-C and Human KHK-A

The intrinsic potency for inhibition of KHK C or A activity may bemeasured using an enzymatic assay which measures the production of FIP.Compounds are prepared in DMSO and tested in a 10-point concentrationcurve, to create 3-fold serial dilutions of the compounds in a 96-wellplate ranging from 20 μM to 1.02 nM. Enzyme is prepared in assay buffer[50 mM 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), 10 mMpotassium chloride, 100 mM magnesium chloride, 2 mMtris(2-carboxyethyl)phosphine (TCEP), 0.01% n-octyl glucoside] andincubated with compounds at RT for 15 min. The reaction is carried outin 100 μL volumes containing substrate concentrations of fructose (250 Mfor KHK-C assay and 1.25 mM for KHK-A assay) and ATP (150 μM for bothisoforms); which are further incubated at RT for 20 min. The reaction isthen halted by the addition of stop buffer; consisting of 0.2% formicacid and 1 μg/ml ¹³C₆-fructose-6-phosphate (¹³C₆-F6P) internal standard.Plates are stored in −20° C. until RapidFire MS analysis.

RapidFire MS Analysis for Quantitation of F1P

An Agilent 300 RapidFire automated extraction system (Agilent, SantaClara, Calif.) with three HPLC quaternary pumps is coupled to an Agilent6495 triple quadrupole mass spectrometer (Agilent Technologies, SantaClara, Calif.) equipped with an electrospray ionization (ESI) interfacesource. The RapidFire Mass Spec system is equipped with a reusableRapidFire C18 (type C) solid-phase extraction (SPE) cartridge (G9205A).

Solvent A, used for sample loading and washing, is 6 mM octylamine(Acros Organics 129495000) brought to pH 5.0 using acetic acid. SolventB, used for sample elution, is 20% water in ACN containing 0.1% formicacid. Samples are sequentially analyzed by aspirating 10 μL onto thecollection loop under vacuum directly from multiwell plates. The 10 μLof sample is loaded onto the C18 cartridge and washed using solvent A ata flow rate of 1.25 mL/min for 5000 ms. The retained analytes are theneluted to the mass spectrometer using solvent B at a flow rate of 1.25mL/min for 5000 ins. The system is re-equilibrated using solvent A at aflow rate of 1.25 mL/min for 2000 is.

The triple quadrupole mass spectrometer is equipped with an ESI sourceand analytes are monitored using selected reaction monitoring (SRM) innegative mode [M−H]−. F1P is monitored at m/z 259.02/96.9 and¹³C₆-fructose-6-phospate is monitored at m/z 264.99/97. The area ratiovalues for F1P is calculated using ¹³C₆-fructose-6-phospate as internalstandard.

The compounds of Examples 1 to 36 were tested essentially as describedabove:

TABLE 6 Example hKHK-C hKHK-A Number IC₅₀ (nM) IC₅₀ (nM)  1 20.7 ± 3.88(n = 4) 24.2 ± 2.9 (n = 4)  2 134 ± 43.1 (n = 7) 98.2 ± 25.0 (n = 7)  341.9 ± 6.77 (n = 3) 45.4 ± 29.5 (n = 4)  4 14.4 ± 1.12 (n = 3) 11.4 ±4.34 (n = 4)  5 57.0 ± 32.6 (n = 5) 37.7 ± 19.9 (n = 5)  6 25.7 ± 5.14(n = 2) 15.4 ± 5.05 (n = 3)  7 18.1 ± 4.85 (n = 2) 6.02 ± 2.27 (n = 3) 8 56.1 ± 62.5 (n = 3) 25.2 ± 15.7 (n = 3)  9 6.26 (n = 1) 3.58 (n = 1)10 418 ± 503 (n = 3) 246 ± 248 (n = 3) 11 8.03 (n = 1) 5.23 (n = 1) 123.87 ± 0.391 (n = 4) 3.65 ± 0.817 (n = 4) 13 15.5 ± 20.4 (n = 3) 7.69 ±7.97 (n = 3) 14 15.9 ± 4.37 (n = 2) 19.1 ± 2.98 (n = 3) 15 27.2 ± 8.60(n = 2) 26.7 ± 3.66 (n = 3) 16 28.0 ± 17.3 (n = 2) 30.0 ± 5.09 (n = 3)17 16.1 ± 7.96 (n = 4) 13.2 ± 12.7 (n = 4) 18 59.2 ± 67.8 (n = 3) 35.6 ±32.4 (n = 3) 19 54.3 (n = 1) 29.2 (n = 1) 20 150 ± 243 (n = 3) 81.8 ±92.6 (n = 3) 21 55.2 ± 28.1 (n = 3) 39.2 ± 13.6 (n = 3) 22 61.3 ± 57.5(n = 3) 33.7 ± 17.8 (n = 3) 23 41.6 ± 47.1 (n = 3) 24.9 ± 15.3 (n = 3)24 7.91 (n = 1) 7.29 (n = 1)  24a 6.06 (n = 1) 9.99 (n = 1)  24b 4.89 (n= 1) 6.75 (n = 1) 25 13.6 ± 7.2 (n = 4) 16.9 ± 9.7 (n = 4) 26 28.4 (n= 1) 22.2 (n = 1) 27 9.21 (n = 1) 4.63 (n = 1) 28 68.0 (n = 1) 51.0 (n= 1) 29 38.6 (n = 1) 103 (n = 1) 30 46.3 (n = 1) 57.8 (n = 1) 31 34.7 (n= 1) 70.5 (n = 1) 32 13.3 (n = 1) 41.5 (n = 1) 33 4.90 ± 1.31 (n = 2)3.63 ± 1.13 (n = 3) 34 3.71 ± 0.178 (n = 2) 5.41 ± 6.13 (n = 3) 35 2.63(n = 1) 1.59 (n = 1) 36 10.0 (n = 1) 3.98 (n = 1) Data expressed asaverage ± SEM (n)

The results as shown in Table 6 above demonstrate that the compounds ofExamples 1 to 36 inhibit the enzymatic activity of both KHIK-C andKHIK-A

KHIK Cellular Activity Assay

Potency can be measured using a cellular assay for the inhibition ofconversion of Fructose to F1P by cellular KHIK. HepG2 liver cells areplated on 96-well cell culture plates in growth media [Dulbecco'sModified Eagle's medium (DMEM) high glucose, 10% heat-inactivated fetalbovine serum (HI FBS), 1× Penicillin/streptomycin] and allowed to attachovernight in a 37° C. incubator. The growth media is washed and replacedwith assay media consisting of Gibco OptiMEM 1 Reduced Serum Medium,0.1% Casein, 8.33 mM D-Fructose-¹³C₆, and compound concentrationsranging from 100 μM to 0.0051 μM (10-point concentration curve). Platesare incubated at 37° C. for 3 h, after which assay media is aspiratedfrom the cell wells. Stop solution consisting of 80% MeOH, 2 mM ammoniumacetate, and 50 ng/mL fructose-6-phosphate-¹³C₆ is then added to thecells. Plates are stored in −20° C. until RapidFire MS analysis(described above).

The compounds of Examples 1 to 36 were tested essentially as describedabove:

TABLE 7 Example HepG2 Number IC₅₀  1 41.1 ± 8.13 (n = 3)  2 98.4 ± 8.69(n = 3)  3 40.2 ± 10.9 (n = 4)  4 16.8 ± 4.97 (n = 4)  5 83.6 ± 8.16 (n= 3)  6 22.9 ± 5.15 (n = 3)  7 8.06 ± 1.41 (n = 3)  8 20.4 ± 8.09 (n =3)  9 15.0 (n = 1) 10 118 ± 24.2 (n = 3) 11 48.4 (n = 1) 12 5.36 ±0.0912 (n = 3) 13 26.2 ± 4.30 (n = 3) 14 34.9 ± 8.26 (n = 3) 15 52.7 ±3.08 (n = 3) 16 73.2 ± 14.9 (n = 3) 17 1.66 ± 0.435 (n = 3) 18 16.3 ±1.84 (n = 3) 19 95.9 (n = 1) 20 35.7 ± 3.47 (n = 3) 21 102 ± 5.47 (n =3) 22 109 ± 9.69 (n = 3) 23 76.2 ± 7.07 (n = 3) 24 27.0 (n = 1)  24a18.2 (n = 1)  24b 27.2 (n = 1) 25 33.5 ± 9.07 (n = 3) 26 81.6 (n = 1) 276.93 (n = 1) 28 144 (n = 1) 29 145 (n = 1) 30 143 (n = 1) 31 119 (n = 1)32 62.9 (n = 1) 33 6.13 ± 0.528 (n = 3) 34 2.93 ± 0.156 (n = 3) 35 6.41(n = 1) 36 13.4 (n = 1) Data expressed as average ± SEM (n)

The results as shown in Table 7 above demonstrate that the compounds ofExamples 1 to 36 inhibit the metabolism of fructose to F1P in HepG2cells.

Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS) Methodfor Pharmacokinetic Assays: Samples are extracted using a proteinprecipitation by adding 180 μL of MeOH:ACN (1:1, v/v) containing aninternal standard to 50 μL of plasma. Samples are then diluted withMeOH:Water (1:1, v/v) to get concentrations within standard curve range.Diluted samples are analyzed by LC-MS/MS using a Sciex API 4000 triplequadrupole mass spectrometer (Applied Biosystems/MDS; Foster City,Calif.) equipped with a TurboIonSpray interface, and operated inpositive ion mode. The analytes are chromatographically separated usinga ECHELON C18 4 um 20×2.1 mm column. LC conditions are Water/1 Mammonium bicarbonate, (2000:10, v/v) (Mobile Phase A), and MeOH/1 Mammonium bicarbonate, (2000:10, v/v) (Mobile Phase B).

Pharmacokinetics in Sprague Dawley Rats

The in vivo pharmacokinetic properties of Example 1 and Example 2 aredemonstrated using Sprague Dawley Rats (fasted; n=3/dose route). Thecompound is administered by a single oral (PO; 2 or 3 mg/kg; volume of10 mL/kg) or intravenous (IV; 1 mg/kg; volume of 1 mL/kg) dose invehicle. Blood is collected from each animal at multiple time pointsbetween 0 and up to 48 hours post-dosage. The plasma concentrations ofExample 1 and Example 2 are determined by a LC-MS/MS method as describedabove.

For Example 1, the mean half-life is 12.9 hours and bioavailability is83% as determined by PO dosing, while IV dosing revealed mean half-lifeis 12.8 hours and the mean clearance is 5.86 mL/min/kg. For Example 2,the mean half-life is 5.12 hours and bioavailability is 95% asdetermined by PO dosing, while IV dosing revealed mean half-life is 4.29hours and the mean clearance is 56.4 mL/min/kg. This data shows Examples1 and 2 have differing levels of clearance, yet both have high oralbioavailability and prolonged elimination evidenced by adequate meanhalf-life.

Pharmacokinetics in Dogs

The in vivo pharmacokinetic properties of Example 1 and Example 2 aredemonstrated using Beagle Dogs (fed, n=3). The compound is administeredby a single oral (PO; 2 or 3 mg/kg; volume of 2 mL/kg) or intravenous(IV; 1 mg/kg; volume of 1 mL/kg) dose in vehicle. Blood is collectedfrom each animal at multiple time points between 0 and up to 72 hourspost-dosage. The plasma concentrations of Example 1 and Example 2 aredetermined by a LC-MS/MS method as described above.

For Example 1, the mean half-life is 36.6 hours and bioavailability is87% as determined by PO dosing, while IV dosing revealed mean half-lifeis 28 hours and the mean clearance is 3.41 mL/min/kg. For Example 2, themean half-life is 9.79 hours and bioavailability is ˜100% as determinedby PO dosing, while IV dosing revealed mean half-life is 10.3 hours andthe mean clearance is 19.6 mL/min/kg. This data shows Examples 1 and 2have differing levels of clearance, yet both have high oralbioavailability and prolonged elimination evidenced by adequate meanhalf-life.

1-17. (canceled)
 18. A method of treating chronic kidney disease in apatient, comprising administering to a patient in need of such treatmentan effective amount of a compound of the formula:

wherein X is N, or C substituted with CN; R¹ is selected from: H

R² and R³ are both H, or one is H and the other is OH; R⁴, R⁵, R⁶, R⁷and R⁹ are independently H or CH₃; R⁸ is H, CH₃, CH₂CH₂OH, C(═O)CH₂NH₂,or C(═O)CH₃; and R¹⁰ is OH or NH₂; or a pharmaceutically acceptable saltthereof.
 19. The method according to claim 18, wherein R¹ is selectedfrom:

or a pharmaceutically acceptable salt thereof.
 20. The method accordingto claim 18, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 21. The method accordingto claim 18, wherein X is N, or a pharmaceutically acceptable saltthereof.
 22. The method according to claim 18, wherein X is Csubstituted with CN, or a pharmaceutically acceptable salt thereof. 23.The method according to claim 18, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 24. The method, accordingto claim 23, wherein the compound is a succinate salt.